中文摘要：

目的 观察富氢液对脂多糖（LPS）诱导的人脐静脉内皮细胞（HUVEC）与单核细胞的黏附及对血管内皮通透性的调节.方法 内皮细胞接种于6孔板,数字表随机分为4组（n=42）：正常对照组（A组）、富氢液组（B组）、LPS组（C组）和LPS＋富氢液组（D组）,B、D组为饱和含氢培养液.待内皮细胞融合成单层后的6、12和24 h,分别加入单核细胞共培养90 min,瑞姬氏染色观察细胞黏附情况,酶联免疫吸附法（ELISA）测定细胞上清液血管细胞黏附分子-1（VCAM-1）和E-选择素的浓度,蛋白印迹法检测血管内皮钙黏连蛋白（VE-cadherin）的表达,免疫荧光法检测24 h时VE-cadherin的分布.结果 与A组比,C组单核-内皮细胞黏附增多（P〈0.05）,E-选择素和VCAM-1浓度显著升高（P〈0.05）,VE-cadherin表达量明显减少（P〈0.05）;相比C组,D组细胞黏附减少（P〈0.05）,VCAM-1和E-选择素的水平下降（P〈0.05）,VE-cadherin表达量增加（P〈0.05）,3个时间点变化趋势相同.24 h荧光结果显示,与A组比,C组VE-cadherin在细胞连接处不完整;与C组比,D组VE-cadherin在细胞连接处较均匀完整.结论 富氢液可减少LPS诱导的黏附分子释放,抑制单核-内皮细胞黏附,并影响VE-cadherin的表达和分布,参与调节血管内皮通透性.

英文摘要：

Objective To explore the regulative effects of hydrogen-rich medium on lipopolysaccharide （LPS）-induced monocytes adhesion to human umbilical vein endothelial cells （HUVEC） and vascular endothelial permeability in vitro. Methods Endothelial cells were seeded in 6-well plates and randomly divided into 4 groups （ n = 42 each ） ： control （ A ）, hydrogen-rich medium （ B ）, LPS （ C ） and LPS ＋ hydrogen-rich medium （ D）. Cells were cultured in plain culture medium in groups A and C or in hydrogen-saturated culture medium in groups B and D. LPS 1 p~g/ml was added into groups C and D. When forming a monolayer, monocytes were added into each group after 6, 12 and 24 h respectively. After a 90- minute co-culturing, adhesion status was detected by Wright-Giemsa stain. Supernatants were collected to detect the concentrations of vascular cell adhesion molecule-1 （ VCAM-1 ） and E-selectin by enzyme-linked immunosorbent assay （ELISA）. The expression of VE-cadherin was measured by Western blot. Cells were stained with immunofluorescence to show the distribution of VE-cadherin after a 24-hour incubation. Results Compared with group A, the adhesion of monocytes to endothelial cells increased （ P 〈 0. 05 ） in group C, the levels of E-selectin and VCAM-1 became elevated （ P 〈 0. 05 ） while the expression of VE-cadherin decreased significantly （ P 〈 0.05 ）. Compared with group C, adhesion decreased in group D （ P 〈 0. 05 ） , the levels of E-selectin and VCAM-1 decreased （ P 〈 0. 05 ） while there was an increased expression of VE- cadherin （ P 〈 0. 05 ）. Three timepoints showed the same tendency. The results of 24 h fluorescence indicated that, compared with group A, VE-cadherin was incomplete in cell-cell connections in group C. However it was complete and well-distributed in group D versus group C. Conclusion Hydrogen-rich medium may reduce the LPS-induced release of adhesion molecules, lessen monocytic adhesion to HUVEC and regulatethe expression of VE-cadh